Enhanced msc preparations

Abstract

The present invention provides preparations of MSCs with important therapeutic potential. The MSC cells are non-primary cells with an antigen profile comprising less than about 1.25% CD45+ cells (or less than about 0.75% CD45+), at least about 95% CD105+ cells, and at least about 95% CD166+ cells. Optionally, MSCs of the present preparations are isogenic and can be expanded ex vivo and cryo-preserved and thawed, yet maintain a stable and uniform phenotype. Methods are taught here of expanding these MSCs to produce a clinical scale therapeutic preparations and medical uses thereof.

1 . A mesenchymal stem cell (MSC) preparation comprising:
 cultured human MSCs expanded from a cryopreserved in-process intermediate MSC preparation;   wherein the cultured human MSCs
 (a) have an antigen profile comprising:
 i. less than 0.75% CD45+ cells; 
 ii. at least 95% CD105+ cells; and 
 iii. at least 95% CD166+ cells; and 
 
 (b) are capable of inhibiting IL2Rα expression by CD3/CD28-activated PBMCs by at least 30% relative to a control. 
   
     
     
         2 - 20 . (canceled) 
     
     
         21 . The MSC preparation of  claim 1 , wherein the MSCs express at least 13 pg TNFR1 per million MSCs. 
     
     
         22 . The MSC preparation of  claim 1 , wherein the MSCs express at least 108 pg TNFR1 per million MSCs. 
     
     
         23 . The MSC preparation of  claim 1 , wherein the MSCs express about 13 μg to about 179 pg TNFR1 per million MSCs. 
     
     
         24 . A frozen cell suspension composition comprising:
 cultured human MSCs expanded from a cryopreserved in-process intermediate MSC preparation, wherein the cultured human MSCs:   (a) have an antigen profile comprising:
 i. less than 0.75% CD45+ cells; 
 ii. at least 95% CD105+ cells; and 
 iii. at least 95% CD166+ cells; and 
   (b) are capable of inhibiting IL2Rα expression by CD3/CD28-activated PBMCs by at least 30% relative to a control;   an electrolyte solution;   at least 5% (v/v) human serum albumin; and   10% DMSO (v/v).   
     
     
         25 . A method of treating steroid refractory acute Graft versus Host Disease (aGvHD) in a pediatric patient comprising intravenously administering four biweekly therapeutic doses of 2×10 6  mesenchymal stem cells/kg. 
     
     
         26 . The method of  claim 25 , wherein the MSCs are allogeneic.